Expression of miR-34a in canine OSA cell lines suppresses cell invasion and migration.

<p>(A) Canine OSA8 and Abrams cells expressing control or pre-miR-34a lentiviral constructs were plated in serum free media in the upper wells of plates for Matrigel invasion assays. The lower chamber of each well contained complete media supplemented with 10% fetal bovine serum. Cells were allowed to invade for 24 hours through a layer of Matrigel basement membrane before removal of Matrigel and media from the upper chambers. Cells that had migrated to the lower surface of the insert membrane were stained with crystal violet and the number of invaded cells were counted in ten random fields in triplicate replicates. Experiments were performed in triplicate and data are represented as the mean ± standard deviation (*<i>p</i> ≤ 0.05, Student’s <i>t</i>-test). (B) Migration behavior was assessed in OSA8 and Abrams cell lines expressing empty vector or pre-miR-34a lentiviral constructs using standard wound-healing assays. Cells were seeded in 6-well plates in complete growth medium and allowed to grow to 70% confluency. A scratch was introduced using a P200 pipet tip and after 24 hours, cells were fixed, stained with crystal violet and evaluated by digital photography.